Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Protein Complex Assembly02:41

Protein Complex Assembly

16.8K
Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
16.8K
Protein Complex Assembly02:41

Protein Complex Assembly

2.6K
2.6K
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

1.8K
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
1.8K
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

6.6K
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
6.6K
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

2.9K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.9K
Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

8.9K
Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electron ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave behind a...
8.9K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Fluorescent Detection of Aqueous <i>N</i>-Nitrosodimethylamine via Photochemical Transformation and Affinity Capture.

Journal of the American Chemical Society·2026
Same author

Boron-Rich Biologics Enabled by Reactive Organic Carboranes.

JACS Au·2026
Same author

High-throughput discovery of arginine-depleted peptides enables effective antisense delivery for Duchenne muscular dystrophy.

bioRxiv : the preprint server for biology·2026
Same author

Expanding the toolkit of LacI/GalR chimeras.

PloS one·2026
Same author

Site-Directed Modification of mRNA with Functionalized Platinum(IV)-Ammines.

JACS Au·2026
Same author

Orthogonal Cleavage of the HMPB Linker from Solid Support Using HFIP.

Organic letters·2025

Related Experiment Video

Updated: Feb 6, 2026

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy
09:30

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy

Published on: August 6, 2018

9.9K

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass

Alexandra J Machen1, Pierce T O'Neil1, Bradley L Pentelute2

  • 1Department of Biochemistry and Molecular Biology, University of Kansas Medical Center.

Journal of Visualized Experiments : Jove
|August 21, 2018
PubMed
Summary

This study demonstrates a novel method for assembling and analyzing anthrax toxin complexes using biolayer interferometry (BLI). The technique allows for real-time monitoring of protein complex dynamics and structural characterization of assembled complexes.

More Related Videos

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
08:07

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry

Published on: January 17, 2025

2.2K
Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

24.8K

Related Experiment Videos

Last Updated: Feb 6, 2026

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy
09:30

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy

Published on: August 6, 2018

9.9K
Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
08:07

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry

Published on: January 17, 2025

2.2K
Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

24.8K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biophysics

Background:

  • Proteins function within macromolecular complexes in vivo.
  • Understanding protein complex assembly and dynamics is crucial for deciphering functional outputs.

Purpose of the Study:

  • To develop and validate a method for monitoring the kinetic assembly and disassembly of protein complexes.
  • To characterize the anthrax toxin pre-endosomal to endosomal complex transition.

Main Methods:

  • Utilized biolayer interferometry (BLI) for label-free monitoring of protein complex formation.
  • Employed disulfide coupling for oriented attachment of lethal factor N-terminal domain (LFN) to a biosensor.
  • Applied electron microscopy (EM) and mass spectrometry (MS) for structural and identity analysis of assembled complexes.

Main Results:

  • Successfully assembled and characterized the anthrax pre-endosomal complex (LFN-PAprepore-CMG2) with high affinity (Kd 170 pM).
  • Demonstrated the transition of the protective antigen (PA) prepore to the PA pore state upon acidification (pH 5.0).
  • Observed weakened binding and dissociation of CMG2 receptor from the PA pore complex.

Conclusions:

  • The developed BLI methodology enables real-time kinetic analysis of protein complex assembly and disassembly.
  • Sequential BLI, EM, and MS provide a robust approach for evaluating protein complex structure and identity.
  • This method facilitates the study of dynamic transitions in biologically relevant macromolecular complexes.